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Novel application of recombinant human urinary trypsin inhibitor

A trypsin inhibition and application technology, applied in the field of recombinant human urinary trypsin inhibitor, can solve the problems of endoplasmic reticulum-related cell apoptosis, tissue and organ damage, etc.

Active Publication Date: 2016-11-23
FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when ERS persists or is too strong, it will cause the occurrence of endoplasmic reticulum-associated apoptosis, which will cause damage to tissues and organs

Method used

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  • Novel application of recombinant human urinary trypsin inhibitor
  • Novel application of recombinant human urinary trypsin inhibitor
  • Novel application of recombinant human urinary trypsin inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Effect of rhUTI on Survival Rate of Mouse Sepsis Model

[0045] 1. Experimental animals and main reagents

[0046] Male BALB / c mice, 6-8w, 20±2g, were purchased from the Institute of Experimental Animals, Chinese Academy of Medical Sciences (animal certificate number: SCXK Beijing 2009-0007). Adaptive feeding for 1 week, free access to food and water, room temperature 25°C, 12h circadian rhythm.

[0047] Anesthetic drugs: Ketamine Hydrochloride Injection (Shanghai First Biochemical Pharmaceutical Company, China), Sumianxin II Injection (developed by Military Veterinary Research Institute, Academy of Military Medical Sciences, China) and 0.9% normal saline (Shandong Luxin Pharmaceutical Group, China) is used after mixing according to the volume ratio of 2:2.5:4.5.

[0048] rhUTI (Shanghai Wanxing Biopharmaceutical Co., Ltd., China): 100,000 U / bottle, ulinastatin for injection (Guangdong Tianpu Biochemical Pharmaceutical Co., Ltd., China): batch number 031211...

Embodiment 2

[0066] Example 2 Effect of rhUTI treatment on the functional status of spleen DC in septic mice

[0067] 1. Experimental animals and main reagents

[0068] The mice and rhUTI used in this example are the same as those in Example 1.

[0069] Mouse CD11c Isolation Kit, Mouse CD4 + T isolation kit: Miltenyi Biltec GmbH, Germany; mouse ELISpot (IL-12, TNF-α) mouse ELISPOT kit (IL-4, IFN-γ): Shanghai Yikesai Biological Products Co., Ltd., China; Anti-mouse CD-86PE, CD-80APC Mouse: Miltenyi Biltec GmbH, Germany; anti-mouse MHC-II, American eBioscience; rabbit anti-mouse GRP78 antibody, American Epitomics; rabbit anti-mouse CHOP antibody, rabbit anti Mouse Caspase-12 was purchased from Cell Signaling Technology, USA.

[0070] 2. Experimental steps:

[0071] (1) The construction of severe sepsis mouse model and the administration of rhUTI were the same as in Example 1.

[0072] (2) After the successful modeling of sepsis, the mice were sacrificed by neck dislocation after 1 day, ...

Embodiment 3

[0080] Example 3 Effect of rhUTI intervention on spleen DC apoptosis in septic mice

[0081] 1. Experimental animals and main reagents

[0082] The mice, rhUTI, and mouse CD11c isolation kits used in this example are the same as those in Example 3.

[0083] Annexin V-PE / 7-AAD Apoptosis Detection Kit (100test, containing 1ml Annexin-V-PE, 500μl 7-AAD and 80ml binding buffer) BD Company, USA.

[0084] 2. Experimental steps:

[0085] ① The construction of severe sepsis mouse model and rhUTI medication are the same as in Example 1.

[0086] ②One day after CLP modeling, they were killed by neck dislocation, and the spleen tissue was collected, minced and ground, and filtered through a 200-mesh steel sieve to separate mononuclear cells from the lymphocyte separation medium. DCs were separated by immunomagnetic beads, and the cell concentration was adjusted. V-PE and 7-AAD were co-incubated at room temperature for 15 min, and DC apoptosis was detected by flow cytometry.

[0087] ...

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Abstract

The invention discloses novel application of a recombinant human urinary trypsin inhibitor. According to the experimental research on animals, the recombinant human urinary trypsin inhibitor has the effects of regulating expression of DC surface molecules CD80, CD86 and MHC-II, influencing T cell proliferation and differentiation, improving maturation state of DC through the expression of surface molecules CD80, CD86 and MHC-II, and further influencing T cell proliferation and differentiation and relieving and regulating negative immunoregulation of T lymphocytes. The recombinant human urinary trypsin inhibitor can achieve the effects of relieving an immunosuppressive state caused by sepsis, particularly cellular immunosuppression, and improving the survival rate of mice suffering from sepsis and the symptoms of sepsis, and can be used for treating sepsis.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a new application of recombinant human urinary trypsin inhibitor. Background technique [0002] Severe burn, trauma, and surgical stress can easily induce infectious complications such as sepsis, and further development can lead to septic shock or multiple organ dysfunction, which is the main cause of death in patients with critical illnesses such as burn, trauma, and surgery. Sepsis has always been a research hotspot at home and abroad because of its high mortality rate and high cost. Although great progress has been made in early fluid resuscitation, antimicrobial drug therapy, nutrition metabolism and auxiliary support of vital organs, the fatality rate of severe sepsis still ranks Not high enough. Therefore, it is undoubtedly of great theoretical and clinical significance to deepen the understanding of sepsis and further explore its possible therapeutic targets. [0003] In recent y...

Claims

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Application Information

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IPC IPC(8): A61K38/57A61P31/00
CPCA61K38/57
Inventor 姚咏明齐安龙祝筱梅孙长海
Owner FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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